Fig. 6: GmLDL2 controls flowering in a GmFER-dependent manner.
a The phenotypes of whole plants and flowers of HX3 and two GmFER-OE transgenic lines under short-day conditions. The two independent GmFER-OE transgenic plants showed early flowering compared to HX3. White arrow, magnified view. Scale bars, 10 cm. b The expression level of GmFER in HX3 and GmFER-OE transgenic plants. GmActin was used as an internal control. c Validation of GmFER expression in GmFER-OE transgenic plants by western blot with anti-Flag antibody. Actin serves as the loading control. d The flowering time of HX3 and GmFER-OE plants under SD (11 h/13 h) and LD (15 h/9 h) conditions. e The design of sgRNAs for CRISPR/Cas9 to knock out GmFER in the Gmldl2 mutant background. sgRNAs targeting CDS are labeled in red. Results of Sanger sequencing of target sites in T1 generations in HX3 and Gmldl2/fer double mutants. sgRNA target and PAM sequences were shown in red and blue, respectively. UTR untranslated region. f Predicted amino acid sequences of GmLDL2 and GmFER in HX3 and Gmldl2/fer double mutants. Missense amino acids resulting from the frameshift caused by CRISPR/Cas9-induced mutations are shown in red. The asterisk indicates the termination of translation resulting from a stop codon. g The phenotypes of whole plants and flowers of HX3, ef1, Gmldl2-2/fer, and Gmldl2-3/fer under long-day conditions. All Gmldl2/fer double mutants showed later flowering compared to ef1. White arrow, magnified view. Scale bars, 10 cm. h The flowering time of all varieties examined under SD (11 h/13 h) and LD (15 h/9 h) conditions. Data represent means ± SD (n = 3 biological replicates) in (b), and data represent means ± SD (n = 10 plants) in (d, h). The P value was generated from a two-sided Student’s t-test. Source data are provided as a Source Data file.
